Purpose: :
Interlamellar separations are frequently seen in corneal histologicalspecimens presumably as a result of fixation. These separations,as well as additional swelling or shrinkage, result in a largevariability in reported central corneal thickness (CCT). Previouslywe reported the use of spectral domain optical coherence tomography(SD-OCT) to obtain in vivo CCT values in C57BL/6 mice. The purposeof the current study was to establish a method of histologicalfixation that produces CCT values similar to in vivo SD-OCTmeasurements and with minimal stromal artifacts.

Conclusions: :
The microwave fixation protocol produced CCT values similarto in vivo measurements and with minimal separation betweenstromal lamellae. The larger CCT measurements from the conventionallyfixed corneas, in the absence of interlamellar separations,were determined to be due to increased stromal thickness, attributedto increased collagen interfibrillar spacing. Interfibrillarspacing in the microwave fixed corneas is very similar to thatreported with X-ray diffraction suggesting microwave fixationpreserves the native collagen structure to a much better degreethan can be obtained by conventional methods.